Mouse denuded oocyte in vitro maturation technology
An in vitro maturation, mouse technology, applied in the field of in vitro maturation of mouse nude eggs
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[0014] 1. Preparation of mature culture medium
[0015] TCM-199 culture medium (Gibco, Grand Island, NY, USA) was supplemented with 1 μg / ml 17β-E 2 , 24.2mg / l sodium pyruvate, 0.05IU / ml FSH, 0.05IU / ml LH and 10ng / ml EGF were used to make mature culture medium. Prepare cysteamine and cystine into concentrated stock solutions of 20mmol / l and 100mmol / l respectively, store at -20°C, and dilute to the required concentration with mature culture medium before use (cysteamine: 100μmol / l; cysteine: 100μmol / l; Acid: 200 μmol / l). Since the commercial TCM-199 culture medium itself contains 83 μmol / l cystine, the final use concentration of cystine is 283 μmol / l.
[0016] 2. Mouse DO preparation
[0017] Ovaries were obtained from superovulated mice, and large follicles were punctured to obtain COC. Use a thin glass tube with a diameter slightly larger than that of oocytes to blow and beat repeatedly to remove cumulus cells to obtain DO.
[0018] 3. Preparation of mouse or goat granulo...
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